Clinical procedure for measuring lipoprotein triglycerides

Abstract

An electrophoresis method of determining the concentration of high density lipoprotein (HDL) triglycerides in fluids, for example, body fluids and simultaneously determining the concentration of very low density lipoprotein (VLDL) and low density lipoprotein (LDL) triglycerides in the fluid. The method includes applying a small sample of the fluid to an electrophoresis support medium, applying a direct current across the medium, applying a developing substrate to the electrophoresed lipoproteins and quantitatively determining the concentration of each lipoprotein triglyceride. This method does permit direct and simultaneous measurement of each lipoprotein triglycerides fraction while eliminating precipitation of each fraction as required by the prior art.

I claim: 
     
       1. A method of simultaneously determining the concentrations of high density lipoprotein triglyceride, very low density lipoprotein triglyceride and low density lipoprotein triglyceride in a sample of body fluid, comprising the steps of: a. applying a small sample of said body fluid to be tested to a solid electrophoresis support media strip,   b. applying a direct current for a predetermined period of time to said support media until the high density, very low density and low density lipoprotein triglycerides have separated on the media,   c. applying a developing substrate sensitive to small concentrations of triglycerides to the electrophoresed lipoprotein strip, and   d. quantitatively determining the concentrations of high density lipoprotein, very low density lipoprotein and low density lipoprotein triglycerides in said body fluid sample from the developed electrophoresed sample.   
     
     
       2. The method of determining concentrations of lipoprotein triglycerides in a sample body fluid defined in claim 1, wherein said developing substrate is an enzymatic triglyceride reagent which is applied to said electrophoresis support media. 
     
     
       3. The method of determining concentrations of lipoprotein triglycerides in a sample of body fluid defined in claim 2, wherein said triglyceride reagent is applied to said support media by immersing the media in a fluid sample of said triglyceride reagent. 
     
     
       4. The method of determining concentrations of lipoprotein triglyceride in a sample of body fluid defined in claim 2, wherein said enzymatic triglyceride substrate is applied to said support media by impregnating an untreated strip of support media with fluid triglyceride reagent and applying said impregnated strip to the electrophoresed lipoprotein triglycerides in a sandwich form and incubating the sandwiched media for a predetermined period of time. 
     
     
       5. The method of determining concentrations of lipoprotein triglyceride in a sample of body fluid defined in claim 1, wherein said electrophoresis support media is cellulose acetate and said direct current is about one hundred eighty (180) volts which is applied to said support media for about twenty minutes. 
     
     
       6. The method of determining concentrations of lipoprotein triglycerides in a sample of body fluid defined in claim 1, wherein the concentrations of the lipoprotein triglycerides are quantitatively determined by a densitometer by measuring absorbance of each lipoprotein triglyceride following application of the developing substrate. 
     
     
       7. The method of determining concentrations of lipoprotein triglyceride in a sample of body fluid defined in claim 1, wherein said quantitative determination is made by eluting each electrophoresed fraction, including high density lipoprotein, very low density lipoprotein and low density lipoprotein triglyceride and then quantitatively determining the concentration of each fraction. 
     
     
       8. The method of determining concentrations of lipoprotein triglycerides in a sample of body fluid defined in claim 7, wherein the concentration of each fraction is determined using a spectrophotometer. 
     
     
       9. The method of determining concentrations of lipoprotein triglycerides in a sample of body fluid defined in claim 7, wherein said triglyceride reagent is tagged with fluorescene, including quantitatively determining the concentration of each fraction by measuring the fluorescence. 
     
     
       10. The method of determining concentrations of lipoprotein cholesterols in a sample of body fluid defined in claim 7, wherein said triglyceride reagent is tagged with a radioactive isotope, including quantitatively determining the concentration of each fraction by measuring the radioactivity of each fraction with a radioisotope counter. 
     
     
       11. A method of determining the concentration of high density lipoprotein triglyceride in body fluid, comprising: a. applying a small sample of said body fluid to a solid electrophoresis support media,   b. applying a direct current across said electrophoresis support media until the high density lipoprotein triglyceride has separated from any remaining lipoprotein in said sample,   c. applying a developing substrate sensitive to high density lipoprotein triglycerides to the separate electrophoresed high density lipoprotein triglyceride, and   d. quantitatively determining the concentration of the high density lipoprotein triglyceride present in said body fluid from said developed electrophoresed sample.   
     
     
       12. The method of determining the concentration of high density lipoprotein triglyceride in a fluid sample defined in claim 11, wherein said developing substrate is an enzymatic triglyceride substrate and said support media is cellulose acetate, including applying said triglyceride reagent to the electrophoresed sample on said cellulose acetate media.